Glyoxal fixation: an immunohistochemistry assay evaluation

Criswell, Sheila; Altman, Savanna R; Peeler, Chelsea; Drake, Taylor M.; Lazar, Cynthia; Douglas, S.; DeJarnatt, Victoria

doi:10.1080/01478885.2021.1996125

Cited by 3 publications

(1 citation statement)

References 10 publications

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“…These results were similar to what already reported for GAF used with human tissues and superior to the suboptimal results obtained with other glyoxal-based fixatives. 12,19,42,69 The protocol to detect the nuclear antigen Ki-67 could be easily optimized with minimal modifications (i.e. increased antigen retrieval) as already suggested by the manufacturer's technical sheets, whereas c-Kit and IBA-1 protocols still need adjustments.…”

Section: Discussionmentioning

confidence: 99%

Non-toxic acid-free glyoxal fixative for veterinary histopathology, immunohistochemistry and molecular analysis

Zappulli

Torrigiani

Detillo³

et al. 2023

Journal of Comparative Pathology

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Section: Discussionmentioning

confidence: 99%

Non-toxic acid-free glyoxal fixative for veterinary histopathology, immunohistochemistry and molecular analysis

Zappulli

Torrigiani

Detillo³

et al. 2023

Journal of Comparative Pathology

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Histochemical and morphological evaluation of a glyoxal acid-free fixative

Peeler,

Pitzer,

Paez

et al. 2024

Biotechnic & Histochemistry

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Non-Toxic Acid-Free Glyoxal Fixative for Veterinary Histopathology, Immunohistochemistry and Molecular Analysis

Zappulli

Torrigiani

Moccia

et al. 2023

Preprint

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Formaldehyde fixation is worldwide the most used system for histopathological examination. However, its toxicity is well known, and preservation of proteins and nucleic acids is not optimal. Alternative fixatives warranting similar morphological quality of tissues and costs, but lacking toxicity and allowing better preservation of proteins and nucleic acids would therefore increase both safety of operators and quality of molecular analysis in pathology. This multi-institutional study aimed to compare the morphological, histochemical, immunohistochemical (IHC), and molecular analyses outcomes of a newly patented, non-toxic, acid-free Glyoxal (GAF) fixative with neutral buffered formaldehyde (NBF). Tissues from a total of 73 subjects were analyzed, including 13 necropsies. Gross features were preserved after GAF fixation, with no tissue hardening or discoloration. Cellular ultrastructure was also better preserved with GAF and histology and histochemistry on GAF-fixed samples showed good results when compared to NBF-fixed samples, with the exception of loss of tinctorial affinity of erythrocytes and mast cell granules. IHC analyses also showed comparable results with only slight and rare protocol adjustment. DNA and RNA yields were higher from GAF-fixed samples (P<0.05) and the tested genes (p53 and COX1) were better amplified. RNA scope showed positive results for c-KIT expression in GAF-fixed mast cell tumors. Based on these data, the non-toxic GAF fixative allows good macroscopical, histological and immunohistochemical analyses of tissue samples, including on-field application, and better molecular analyses when compared to NBF. This represents a promising possibility for teaching, diagnostic, and research in veterinary pathology.

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Glyoxal fixation: an immunohistochemistry assay evaluation

Cited by 3 publications

References 10 publications

Non-toxic acid-free glyoxal fixative for veterinary histopathology, immunohistochemistry and molecular analysis

Non-toxic acid-free glyoxal fixative for veterinary histopathology, immunohistochemistry and molecular analysis

Histochemical and morphological evaluation of a glyoxal acid-free fixative

Non-Toxic Acid-Free Glyoxal Fixative for Veterinary Histopathology, Immunohistochemistry and Molecular Analysis

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